Pertussis, also known as whooping cough, is the only vaccine-preventable infection that remains endemic in the U.S. Moreover, more than 48, 277 cases were reported to Centers for Diseases Control and Prevention (CDC) in 2012, the highest since 1959. Pertussis is caused by the bacterium Bordetella pertussis (B. pertussis). The disease is highly contagious and can be difficult to diagnose. Most of the cases present a mild or subclinical infection. Pertussis vaccine-induced immunity is not permanent thus the immunized host is vulnerable to re- infection later in life. The purpose of this application is to develop a low-cost, instrument-free paper-based point-of-care (POC) biochip for rapid diagnosis of pertussis in limited-resource settings. Our central hypothesis is that the integration of paper-based microfluidic technology with specific DNA testing can provide a fast, cost-effective and accurate diagnosis that can be used in various venues. The proposed methodology will allow for the confirmation of suspected cases of pertussis at medical offices, schools, and in the field. To accomplish our goal we have developed these specific aims: (1) Develop a low-cost paper-based microfluidic point-of-care device for the instrument-free detection of B. pertussis; (2) Validate the paper-based device by testing clinical samples. The long-term goals of this research are to design and validate a paper-based biochip for the identification of a wide variety of infectious diseases.